Encoded by the CPE gene and also known as Carboxypeptidase H, CPE is a single chain peptidase with an optimal pH range between 5.0‑6.0. It is a zinc metallocarboxypeptidase that removes basic amino acids from the C-terminus of peptides (1). Like other metallocarboxypeptidases, its activity is stimulated by millimolar concentrations of Co2+. Its activity is regulated by pH-induced aggregation above pH 6.0. Its major function seems to process numerous peptide hormones and neurotransmitters. In addition to its proteolytic function, it also plays a role as a sorting receptor (2), which may be attributed to the sorting of this protein into the secretory pathway. The C-terminal domain of CPE causes the peripheral association of CPE with membranes below neutral pH, resulting in the association of this protein into membranes (3). CPE knockout mice live but become obese due to impaired glucose clearance and insulin resistance (4).
Recombinant Human Carboxypeptidase E/CPE Protein, CF
R&D Systems | Catalog # 3587-ZN
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Key Product Details
- R&D Systems NS0-derived Recombinant Human Carboxypeptidase E/CPE Protein (3587-ZN)
- Quality control testing to verify active proteins with lot specific assays by in-house scientists
- All R&D Systems proteins are covered with a 100% guarantee
Source
NS0
Accession Number
Structure / Form
Mature form
Applications
Enzyme Activity
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Product Specifications
Source
Mouse myeloma cell line, NS0-derived human Carboxypeptidase E/CPE protein
Arg42-Ser453, with a C-terminal 6-His tag
Arg42-Ser453, with a C-terminal 6-His tag
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
Arg42
Predicted Molecular Mass
47 kDa
SDS-PAGE
57 kDa, reducing conditions
Activity
Measured by its ability to cleave a peptide substrate, benzoyl-AR-OH. The product, Arg, is reacted with ortho-phthaldialdehyde (OPA) to form a fluorescent molecule.
The specific activity is >12,000 pmol/min/µg, as measured under the described conditions.
The specific activity is >12,000 pmol/min/µg, as measured under the described conditions.
Formulation, Preparation, and Storage
3587-ZN
| Formulation | Supplied as a 0.2 μm filtered solution in MES and NaCl. |
| Shipping | The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
Background: Carboxypeptidase E/CPE
References
- Fricker, L.D. (2004) in Handbook of Proteolytic Enzymes, Barrett, A.J. et al. eds., pp. 840.
- Cool, D.R. et al. (1997) Cell 88:73.
- Zhang, C-F. et al. (2003) Biochem. J. 369:453.
- Cawley, N.X. et al. (2004) Endocrinology 145:5807.
Alternate Names
CPE, CPH-1
Gene Symbol
CPE
UniProt
Additional Carboxypeptidase E/CPE Products
Product Documents for Recombinant Human Carboxypeptidase E/CPE Protein, CF
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Human Carboxypeptidase E/CPE Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Human Carboxypeptidase E/CPE Protein, CF
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Protocols
View specific protocols for Recombinant Human Carboxypeptidase E/CPE Protein, CF (3587-ZN):
Materials
- Assay Buffer: 50 mM NaOAc, 5 µM ZnCl2, pH 5.5
- Recombinant Human Carboxypeptidase E/CPE (rhCPE) (Catalog # 3587-ZN)
- Substrate: Benzoyl-Ala-Arg-OH (Bachem, Catalog # G-4145), 50 mM stock in DMSO
- ortho-phthaldialdehyde (OPA) (134.13 g/mol) (Sigma, Catalog # P0657), 50 mg/mL stock in DMSO
- OPA Buffer: 0.2 M NaOH containing 0.1% mercaptoethanol (v/v)
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rhCPE to 0.1 ng/µL in Assay Buffer.
- Dilute Substrate to 1 mM in Assay Buffer.
- Combine 75 µL of 0.1 ng/μL rhCPE and 75 µL of 1 mM Substrate. Include a Blank containing 75 µL of Substrate only.
- Incubate for 10 minutes at room temperature.
- Add 150 µL of 15 mM OPA in 0.2 M NaOH containing 0.1% mercaptoethanol (v/v) to reaction vials and the Blank (stops reaction).
- After stopping the reaction, add 75 µL of 0.1 ng/µL rhCPE to the Blank.
- Mix well and incubate at room temperature for 2-10 minutes.
- Load 200 µL of sample(s) and blank into plate.
- Read at excitation and emission wavelengths of 330 nm and 450 nm (top read), respectively in endpoint mode.
- Calculate specific activity:
|
Specific Activity (pmol/min/µg) = |
Adjusted Fluorescence* (RFU) x Conversion Factor** (pmol/RFU) |
| Incubation time (min) x amount of enzyme (µg) |
*Adjusted for Substrate Blank
**Derived using calibration standard L-Arginine (Sigma, Catalog # A-5006).
- rhCPE: 0.005 µg
- Substrate: 0.25 mM
- OPA: 7.5 mM
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